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    • 99. 发明申请
    • METHOD FOR GENERATING CLONED ANIMALS USING CHROMOSOME SHUFFLING
    • 使用染色体沉淀法生成克隆动物的方法
    • WO2002077175A2
    • 2002-10-03
    • PCT/US2002/008842
    • 2002-03-25
    • ADVANCED CELL TECHNOLOGY, INC.WEST, Michael, D.CIBELLI, Jose
    • WEST, Michael, D.CIBELLI, Jose
    • C12N
    • A01K67/0275A01K2217/05A01K2227/101C12N15/873C12N15/877C12N15/8771C12N2517/00C12N2517/04
    • The present invention concerns the use of chromosomal replacement techniques in the context of producing cloned and transgenic animals, in order to correct chromosome abnormalities or alter autosomal genotypes, and provide for novel breeding pairs by replacing the sex chromosome in animals to be cloned. Replacement of a sex chromosome, or an X or Y chromosome, will result in animals that are autosomally isogenic and sexually non-isogenic (AISN), with ''autosomally isogenic'' meaning that the paired sets of autosomes (non-sex chromosomes) in each animal are isogenic or identical. Also included in the invention are animals that are both ''autosomally'' and ''allelically'' isogenic whereby each particular pair of chromosomes is internally isogenic or identical within a single animal as well as between animals. Such animals are particularly useful in generating a line of cloned mammals using sexual reproduction, without having to undergo nuclear transfer in order to propagate cloned animals.
    • 本发明涉及在产生克隆和转基因动物的背景下使用染色体替代技术,以便校正染色体异常或改变常染色体基因型,并通过替代待克隆的动物中的性染色体来提供新的育种对。 更换性染色体或X或Y染色体将导致常染色体同源异体和性别非同基因(AISN)的动物,具有“常染色体同源基因”,这意味着成对的常染色体组(非性染色体) 在每只动物中都是同基因的或相同的。 本发明还包括既“常染色体”和“等位性”同源的动物,由此每一个特定的一对染色体在单个动物内部以及动物之间内部是同基因的或相同的。 这样的动物特别可用于使用有性繁殖产生一系列克隆的哺乳动物,而不必经受核转移以繁殖克隆的动物。
    • 100. 发明申请
    • METHODS FOR ASSAYING GENE IMPRINTING AND METHYLATED CpG ISLANDS
    • 用于测定基因印迹和甲基化CpG岛的方法
    • WO0190313A3
    • 2002-05-16
    • PCT/US0116253
    • 2001-05-22
    • UNIV JOHNS HOPKINSFEINBERG ANDREWSTRICHMAN ALMASHANU LIORAJIANG SHAN
    • FEINBERG ANDREWSTRICHMAN-ALMASHANU LIORAJIANG SHAN
    • A01K67/027A61P35/00C12N5/074C12N5/10C12N15/09C12Q1/02C12Q1/68C12N5/06C12N15/11G01N33/50
    • C12N5/0611C12N2501/115C12N2501/125C12N2501/235C12N2501/385C12N2502/13C12N2503/00C12N2503/02C12N2510/00C12N2517/00C12Q1/6827C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/154
    • Genomic imprinting is a parent of origin-dependent gene silencing that involves marking of alleles in the germline and differential expression in somatic cells of the offspring. Imprinted genes and abnormal imprinting have been implicated in development, human disease, and embryonic stem cell transplantation. We have established a model system for genomic imprinting using pluripotent 8.5 d.p.c. mouse embryonic germ (EG) cell lines derived from an interspecific cross. We find that allele-specific imprinted gene expression has been lost in these cells. However, partial restoration of allele-specific silencing can occur for some imprinted genes after in vitro differentiation of EG cells into somatic cell lineages, indicating the presence of a gametic memory that is separable from allele-specific gene silencing. We have also generated a library containing most methylated CpG islands. A subset of these clones was analyzed and revealed a subdivision of methylated CpG islands into 4 distinct subtypes: CpG islands belonging to high copy number repeat families; unique CpG islands methylated in all tissues; unique methylated CpG islands that are unmethylated in the paternal germline; and unique CpG islands methylated in tumors. This approach identifies a methylome of methylated CpG islands throughout the genome.
    • 基因印记是起源依赖基因沉默的亲本,其涉及在种系中标记等位基因和在后代的体细胞中的差异表达。 印迹基因和异常印记涉及发育,人类疾病和胚胎干细胞移植。 我们已经建立了使用多能8.5 d.p.c的基因组印迹模型系统。 来自种间交叉的小鼠胚胎胚芽(EG)细胞系。 我们发现在这些细胞中已经丢失了等位基因特异性印记的基因表达。 然而,在体外将EG细胞分化为体细胞谱系后,一些印迹基因可能部分恢复等位基因特异性沉默,表明存在可与等位基因特异性基因沉默分离的配子记忆 。 我们还生成了一个包含大多数甲基化CpG岛的文库。 分析了这些克隆的一个子集,并揭示了甲基化CpG岛分为4个不同的亚型:属于高拷贝数重复家族的CpG岛; 所有组织中独特的CpG岛甲基化; 在父系种系中未甲基化的独特的甲基化CpG岛; 和独特的CpG岛在肿瘤中甲基化。 该方法鉴定整个基因组中的甲基化CpG岛的甲基化。